FIG. 4.

Deletion of YNG2 affects NuA4 activity and transcription of NuA4 target genes. (A) Protein extracts from deleted strain QY203 (Δyng2) or QY203, transformed with pAN104, and expressing HA-Yng2p (WT) were fractionated on a MonoQ column. Fractions were assayed for HAT activity and Western blotted for the indicated NuA4 components. (B) MonoQ NuA4 peak fractions of wild type (WT) and Δyng2, shown in panel A, were pooled and loaded on a Superose-6 gel filtration column. Eluted fractions were tested for the presence of NuA4 components and Ada2p by Western blot. HAT assays with nucleosomes and free histones as substrates were also performed. (C) The strains QY204 (WT) and QY203 (Δyng2) were grown in YPD to an OD of 1.0. Total RNA was extracted and Northern blots were done, using PHO5, HIS4, and ACT1 probes. The deletion of YNG2 significantly affected the transcript levels of PHO5 and HIS4 but not ACT1.